Disease symptoms of some phytopathogenic fungi are associated with changes in cytokinin (CK) levels. Here, we show that the CK profile of ergot-infected rye plants is also altered, although no pronounced changes occur in the expression of the host plant's CK biosynthesis genes. Instead, we demonstrate a clearly different mechanism: we report on the first fungal de novo CK biosynthesis genes, prove their functions and constitute a biosynthetic pathway. The ergot fungus Claviceps purpurea produces substantial quantities of CKs in culture and, like plants, expresses enzymes containing the isopentenyltransferase and lonely guy domains necessary for de novo isopentenyladenine production. Uniquely, two of these domains are combined in one bifunctional enzyme, CpIPT-LOG, depicting a novel and potent mechanism for CK production. The fungus also forms trans-zeatin, a reaction catalysed by a CK-specific cytochrome P450 monooxygenase, which is encoded by cpp450 forming a small cluster with cpipt-log. Deletion of cpipt-log and cpp450 did not affect virulence of the fungus, but Δcpp450 mutants exhibit a hyper-sporulating phenotype, implying that CKs are environmental factors influencing fungal development.

• MeSH
• alkyltransferasy a aryltransferasy metabolismus   MeSH
• Claviceps genetika   růst a vývoj   metabolismus   MeSH
• cytokininy biosyntéza   MeSH
• delece genu MeSH
• geny hub genetika   MeSH
• isopentenyladenosin biosyntéza   MeSH
• rostlinné geny genetika   MeSH
• systém (enzymů) cytochromů P-450 genetika   MeSH
• žito mikrobiologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adenylate isopentenyltransferase MeSH Prohlížeč
• alkyltransferasy a aryltransferasy MeSH
• cytokininy MeSH
• isopentenyladenosin MeSH
• N(6)-(delta(2)-isopentenyl)adenine MeSH Prohlížeč
• systém (enzymů) cytochromů P-450 MeSH

Claviceps purpurea, C. grohii, C. zizaniae, C. cyperi, and C. nigricans are closely related ergot fungi and form a monophyletic clade inside the genus Claviceps. Analysis of alkaloid content in C. nigricans sclerotia using UPLC detected ergocristine (1), ergosine (2), alpha-ergocryptine (3), and ergocristam (4). Alkaloids 1, 3, and 4 were found in the sclerotia of C. grohii. The content of 4 in the mixture of alkaloids from C. nigricans and C. grohii (over 8% and over 20%, respectively) was unusually high. Submerged shaken cultures of C. nigricans produced no alkaloids, whereas C. grohii culture formed small amounts (15 mg L (-1)) of extracellular clavines and 1. In the previously used HPLC method the ergocristam degradation product could have been obscured by the ergosine peak. Therefore sclerotia of a C. purpurea habitat-specific population G2 with the dominant production of 1 and 2 have been reanalyzed, but no 4 was detected. The phylogeny of the C. purpurea-related species group is discussed with regard to alkaloid-specific nonribosomal peptide synthetase duplication leading to the production of two main ergopeptines instead of a single product.

• MeSH
• alkaloidy chemie   klasifikace   izolace a purifikace   MeSH
• Claviceps chemie   genetika   růst a vývoj   MeSH
• DNA analýza   MeSH
• molekulární struktura MeSH
• peptidsynthasy metabolismus   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• alkaloidy MeSH
• DNA MeSH
• non-ribosomal peptide synthase MeSH Prohlížeč
• peptidsynthasy MeSH

Oligosaccharides produced by submerged cultures of C. africana and C. sorghi were isolated by semipreparative HPLC. Structure of 6-O-beta-D-fructofuranosyl-D-glucopyranose (blastose), 1,6-bis-O-(beta-D-fructofuranosyl)-alpha-D-glucopyranoside (neokestose) and two sugar alcohols, 1-O-beta-D-fructofuranosyl-D-mannitol (fructosylmannitol) and 1,6-bis-O-(beta-D-fructofuranosyl)-D-mannitol (bisfructosylmannitol) was determined by NMR spectrometry. MALDI TOF MS analysis revealed molecular ions [M+Na]+ that indicate the presence of other tetra- and pentasaccharides (m/z = 689.4 and 851.5, respectively) and corresponding sugar alcohol (m/z = 691.4). Rapid conversion of sucrose into series of oligosaccharides and corresponding sugar alcohols was observed in all tested strains.

• MeSH
• Claviceps růst a vývoj   izolace a purifikace   metabolismus   patogenita   MeSH
• cukerné alkoholy chemie   metabolismus   MeSH
• fermentace MeSH
• molekulární struktura MeSH
• nemoci rostlin mikrobiologie   MeSH
• nukleární magnetická rezonance biomolekulární MeSH
• oligosacharidy biosyntéza   chemie   izolace a purifikace   MeSH
• Sorghum mikrobiologie   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cukerné alkoholy MeSH
• oligosacharidy MeSH

Types of asexual sporulation in 17 Claviceps species and the closely related Corallocytostroma ornicopreoides were revised in relation to the phylogeny of clavicipitaceous fungi. We observed: (1) enteroblastic conidiation from branched phialidic conidiophores typical of the genus (anamorph Sphacelia) in all species including Corallocytostroma; (2) widespread and often sequential formation of terminal holoblastic secondary conidia on tapering hyphae arising from sphacelial macroconidia: and (3) in addition to sphacelial conidiation, sympodial holoblastic conidiation of the Ephelis-type in cultures of C. zizaniae and in both the culture and sphacelial tissue of C. citrina. Secondary conidiation was not found in C. purpurea, C. citrina and C. sorghicola. During sphacelial fructification, most species produced macroconidia and microconidia. Only macroconidia formed in planta underwent secondary conidiation whereas microconidia did not germinate at all. In C. phalaridis, the formation of holoblastic 2-3 celled appendaged conidia was observed, similar to that of Aciculosporium and Neoclaviceps. In dendrograms based on ITS1-5.8S rDNA-ITS2 sequences, genera and species with appendaged conidia grouped on a highly supported clade with ancestral Corallocytostroma. The clade was placed inside a group of tropical species of Claviceps, without any relationship to Balansiae.

• MeSH
• Claviceps klasifikace   růst a vývoj   fyziologie   MeSH
• DNA fungální analýza   MeSH
• fylogeneze * MeSH
• Hypocreales klasifikace   růst a vývoj   fyziologie   MeSH
• kultivační média MeSH
• lipnicovité mikrobiologie   MeSH
• mezerníky ribozomální DNA analýza   MeSH
• ribozomální DNA analýza   MeSH
• RNA ribozomální 5.8S genetika   MeSH
• spory hub fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA fungální MeSH
• kultivační média MeSH
• mezerníky ribozomální DNA MeSH
• ribozomální DNA MeSH
• RNA ribozomální 5.8S MeSH

Among 18 tested strains of Claviceps spp., 7 produced significant amounts of exocellular polysaccharide (EPS). The maximum production of EPS was found in fermentation broth of Claviceps viridis. The kinetics of growth, substrate consumption, and EPS production in the batch, aerobic, submerged culture of this fungus were investigated in detail. The experimental data were processed by a simple mathematical model describing mass balance of growth, substrate consumption, formation of intermediates, and production of EPS. The parameters of the model were estimated from data obtained in cultivation performed in flasks and two laboratory fermentors of different size. Physiological similarity was obtained during process scale-up in volumetric ratio 1:100. The sugar consumption efficiency (52%) and observed EPS productivity (1.9 kg/m3 per d) were comparable with literature data.

• MeSH
• biologické modely MeSH
• biomasa MeSH
• Claviceps růst a vývoj   metabolismus   MeSH
• fermentace MeSH
• glukany biosyntéza   MeSH
• kinetika MeSH
• kultivační média MeSH
• kyslík metabolismus   MeSH
• rozpustnost MeSH
• sacharosa metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glukany MeSH
• kultivační média MeSH
• kyslík MeSH
• sacharosa MeSH

A non-mutant saprophytic strain C. paspali which forms conidia both on a solid medium and during submerged fermentation is described. Conidiation proceeded in parallel with culture growth and production of alkaloids. The effect of composition of culture media on the intensity of conidiation is described.

• MeSH
• časové faktory MeSH
• Claviceps růst a vývoj   metabolismus   fyziologie   MeSH
• ergoliny biosyntéza   MeSH
• kultivační média MeSH
• kyselina lysergová biosyntéza   fyziologie   MeSH
• spory fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ergoliny MeSH
• kultivační média MeSH
• kyselina lysergová MeSH

Clomiphene depressed the growth and enhanced clavine production of Claviceps purpurea strains 129,35 and 59. Mycelial content of 18:2 and 16:0 fatty acids decreased, whereas that of 18:1 and 18:0 acids increased. In the mutant strain 59 clomiphene, triadimefon and ergosterol stimulated the impaired function of chanoclavine cyclase. Their effect was counteracted by plant oil. Clomiphene decreased the content of total lipids (44%), triglycerides (32%), sterols (22%) and sterol/phospholipid molar ratio. The PC/PE ratio was 9X increased. Clomiphene and triadimefon enhanced membrane fluidity of protoplasts, ergosterol and oil reverted their effect.

• MeSH
• Claviceps účinky léků   růst a vývoj   metabolismus   MeSH
• fluidita membrány účinky léků   MeSH
• fosfolipidy metabolismus   MeSH
• kinetika MeSH
• klomifen farmakologie   MeSH
• mastné kyseliny metabolismus   MeSH
• metabolismus lipidů MeSH
• steroly metabolismus   MeSH
• triglyceridy metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fosfolipidy MeSH
• klomifen MeSH
• mastné kyseliny MeSH
• steroly MeSH
• triglyceridy MeSH

The proposed method includes primary and secondary treatments. Of the various disinfectants tested, 1-propanol and 1-butanol as a primary treatment and phenol, resorcinol, HgC12 or NaCl as a secondary treatment completely eliminated the contamination hazard. 1-Butanol and phenol were not useful disinfectants since they inhibited the growth of C. fusiformis along with the other contaminants. Primary and secondary treatment of sclerotia with 1-propanol and resorcinol, respectively, produced the maximum of stable cultures. Out of 120 cultures tested, 5 cultures demonstrated an appreciable yield of alkaloids under submerged cultural conditions.

• MeSH
• 1-propanol farmakologie   MeSH
• Claviceps růst a vývoj   izolace a purifikace   metabolismus   MeSH
• dezinficiencia farmakologie   MeSH
• metody MeSH
• námelové alkaloidy biosyntéza   MeSH
• resorcinoly farmakologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 1-propanol MeSH
• dezinficiencia MeSH
• námelové alkaloidy MeSH
• resorcinoly MeSH

Panicum repens and P. antidotale were found to be infected with Claviceps sp. This is the first report of ergot on P. repens. The pyrenomycete produced abundant sclerotia on the host plants. The sclerotia contained 0.71 and 0.68% alkaloids, respectively, which predominantly consisted of chanoclavine, festuclavine and agroclavine. The infected grasses were possibly mycotoxic. Submerged cultures of Claviceps strain isolated from Panicum spp. produced significant amount of chanoclavine, festuclavine and agroclavine. No pharmaceutically important alkaloid was found in sclerotia or in submerged culture.

• MeSH
• Claviceps růst a vývoj   metabolismus   MeSH
• lipnicovité mikrobiologie   MeSH
• námelové alkaloidy biosyntéza   MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Indie MeSH
• Názvy látek
• námelové alkaloidy MeSH

Claviceps paspali FA produced high concentrations of alkaloid under submerged conditions. Their production was found to depend on the developmental stage and treatment of the filamentous culture inoculum. A medium containing Bacto-peptone with a constant composition of amino acids was selected for the preparation of the inoculum. A two-week fermentation in a synthetic medium with mannitol at 24 +/- 1 degrees C resulted in an increased production of total alkaloids from the original value of 100-200 micrograms/mL to more than 2000 micrograms/mL. Addition of tryptophan did not further increase the production of alkaloids but resulted in changes of the spectrum of some metabolites, 2,3-Dihydroxybenzoic acid accompanied the alkaloids in the fermentation medium. alpha-Hydroxyethyllysergamide was the predominant component of extracellular alkaloids (80% in the first days of fermentation). During fermentation the level of this alkaloid continuously decreased while the concentration of the accompanying alkaloids, i.e. lysergamide and the corresponding minor isomers, increased.

• MeSH
• alkaloidy biosyntéza   MeSH
• Claviceps růst a vývoj   metabolismus   MeSH
• fermentace MeSH
• glukosa metabolismus   MeSH
• kultivační média MeSH
• kyselina lysergová analogy a deriváty   biosyntéza   MeSH
• mannitol metabolismus   MeSH
• tryptofan farmakologie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alkaloidy MeSH
• glukosa MeSH
• kultivační média MeSH
• kyselina lysergová MeSH
• mannitol MeSH
• tryptofan MeSH